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ab140359 is an In-Cell ELISA (ICE) assay kit that uses quantitative immunocytochemistry to measure levels of COX1 and SDHA levels in cultured cells. Cells are fixed in a microplate and targets of interest are detected with highly specific, well-characterized antibodies. Relative target levels are quantified using secondary antibodies conjugated to either horseradish peroxidase (HRP) or alkaline phosphatase (AP) which generate signal through the use of two spectrally distinct fluorogenic substrates. Fluorescence is measured using a standard fluorescent spectrophotometer and relative levels of target proteins are quantified. Optionally, antibody signal intensity can be normalized to the total cell amount using Janus Green stain. In-Cell ELISA (ICE) technique generates quantitative data with specificity similar to Western blotting, but with much greater quantitative precision and higher throughput due to the greater dynamic range and linearity of fluorescence detection and the ability to run up to 96 samples in parallel. This method rapidly fixes the cells in situ, stabilizing the in vivo levels of proteins and their post-translational modifications, and thus essentially eliminates changes during sample handling, such as preparation of protein extracts.
Plates are available in our ICE (In-Cell ELISA) Support Pack (ab111542) which can be bought seperately.
"Cytochrome c oxidase is the component of the respiratory chain that catalyzes the reduction of oxygen to water. Subunits 1-3 form the functional core of the enzyme complex. CO I is the catalytic subunit of the enzyme. Electrons originating in cytochrome c are transferred via the copper A center of subunit 2 and heme A of subunit 1 to the bimetallic center formed by heme A3 and copper B. Catalytic activity carries out the following reaction;
4 ferrocytochrome c + O2 + 4 H+ = 4 ferricytochrome c + 2 H2O.
Flavoprotein (FP) subunit of succinate dehydrogenase (SDH) that is involved in complex II of the mitochondrial electron transport chain and is responsible for transferring electrons from succinate to ubiquinone (coenzyme Q). Can act as a tumor suppressor. Catalytic activity carries out following reaction;
Succinate + ubiquinone = fumarate + ubiquinol.
|Components||Identifier||1 x 96 tests|
|1000X Anti-Mouse IgG/AP-Labeled Secondary Antibody||1 x 20µl|
|1000X HRP Labeled Secondary Antibody (Anti-Mouse IgG2a)||1 x 20µl|
|100X Primary Antibody Cocktail||1 x 120µl|
|100X Triton X-100||1 x 1.25ml|
|10X Blocking Buffer||1 x 15ml|
|10x Phosphate buffered Saline (PBS)||8209706||1 x 100ml|
|10X Quenching Solution||1 x 1.5ml|
|400X Fluorescent Substrate Cocktail||1 x 50µl|
|400X Tween-20||1 x 2ml|
|8000X Hydrogen Peroxide||1 x 50µl|
|Fluorescent Substrate Buffer||1 x 12ml|
|Janus Green Stain||1 x 17ml|
Our Abpromise guarantee covers the use of ab140359 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|In-Cell ELISA||Use at an assay dependent concentration.|
ab140359 has not yet been referenced specifically in any publications.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"